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14 results on '"MESH : Stem Cells"'

1. Comparative Study of Injury Models for Studying Muscle Regeneration in Mice

Author

Shahragim Tajbakhsh, Cédric Thépenier, David Briand, Aurélie Guguin, Aurore Besnard, Mathilde Latil, Pierre Rocheteau, Grégory Jouvion, Jean-Marc Cavaillon, Quentin Pascal, Fabrice Chrétien, David Hardy, Barbara Gayraud-Morel, Histopathologie humaine et Modèles animaux, Institut Pasteur [Paris], Université Paris Descartes - Paris 5 ( UPD5 ), Institut de Recherche Biomédicale des Armées ( IRBA ), Plateforme de Cytométrie en Flux, GHU A. Chenevier - Henri Mondor ( Inserm U955, équipe 16 ), Cellules Souches et Développement, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Cytokines et Inflammation, This work was financially supported by AFM-vaincre les myopathies ([http://www.afm-telethon.fr/] Role: design, data collection and analysis), Fondation ³Les gueules cassées², Institut Pasteur and Région Ile de France. his work wasalso supported by the agence-nationale-recherche ([http://www.agence-nationale-recherche.fr/], Role:design, data collection and analysis). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of themanuscript, Institut Pasteur [Paris] (IP), Université Paris Descartes - Paris 5 (UPD5), Institut de Recherche Biomédicale des Armées (IRBA), GHU A. Chenevier - Henri Mondor (Inserm U955, équipe 16), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), BENEDIC, Bénédicte, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut de Recherche Biomédicale des Armées [Brétigny-sur-Orge] (IRBA)

Subjects
Pathology, MESH : Cytokines, Barium Compounds, MESH: Barium Compounds, Pathology and Laboratory Medicine, Muscle Development, MESH : Green Fluorescent Proteins, Mice, Animal Cells, Fibrosis, Freezing, MESH : Chlorides, MESH: Animals, lcsh:Science, MESH : Muscle, Skeletal, Immune Response, Musculoskeletal System, MESH : Macrophages, Innate Immune System, MESH: Cytokines, MESH : Neovascularization, Physiologic, Stem Cells, MESH: Satellite Cells, Skeletal Muscle, MESH : Mice, Transgenic, 3. Good health, [SDV] Life Sciences [q-bio], Neurology, MESH: Cold Injury, Cytokines, Cellular Types, Muscle Regeneration, medicine.medical_specialty, Satellite Cells, Skeletal Muscle, Immune Cells, Immunology, MESH : Muscle Development, Neovascularization, Physiologic, MESH : Myoblasts, Necrosis, 03 medical and health sciences, Signs and Symptoms, Cardiotoxin, MESH: Green Fluorescent Proteins, Regeneration, MESH: Chlorides, Cold Injury, MESH: Elapid Venoms, MESH : Cobra Cardiotoxin Proteins, Blood Cells, MESH: Vascular Endothelial Growth Factor Receptor-2, [ SDV ] Life Sciences [q-bio], [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, Macrophages, Regeneration (biology), lcsh:R, Biology and Life Sciences, Molecular Development, medicine.disease, Vascular Endothelial Growth Factor Receptor-2, MESH : Necrosis, Mice, Inbred C57BL, Biological Tissue, 030104 developmental biology, Skeletal Muscles, lcsh:Q, Organism Development, Developmental Biology, MESH: Freezing, 0301 basic medicine, Physiology, [SDV]Life Sciences [q-bio], Cobra Cardiotoxin Proteins, Poison control, lcsh:Medicine, Myoblasts, White Blood Cells, Immune Physiology, Neurobiology of Disease and Regeneration, Morphogenesis, Medicine and Health Sciences, Myocyte, MESH: Muscle, Skeletal, Multidisciplinary, Muscles, MESH: Regeneration, MESH: Cobra Cardiotoxin Proteins, medicine.anatomical_structure, MESH: Models, Animal, Models, Animal, MESH: Fibrosis, MESH : Barium Compounds, MESH : Stem Cells, MESH : Fibrosis, MESH: Muscle Development, Basal lamina, Anatomy, Stem cell, medicine.symptom, MESH: Neovascularization, Physiologic, Research Article, MESH : Cold Injury, Histology, MESH: Mice, Transgenic, MESH : Elapid Venoms, Green Fluorescent Proteins, MESH : Vascular Endothelial Growth Factor Receptor-2, Muscle Tissue, Mice, Transgenic, MESH : Regeneration, MESH: Stem Cells, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH : Mice, Inbred C57BL, Biology, Andrology, Chlorides, MESH: Mice, Inbred C57BL, MESH : Satellite Cells, Skeletal Muscle, MESH : Mice, medicine, Animals, MESH: Myoblasts, Muscle, Skeletal, MESH: Mice, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Inflammation, Elapid Venoms, MESH: Necrosis, MESH: Macrophages, Cell Biology, MESH : Models, Animal, MESH : Freezing, Immune System, MESH : Animals
Abstract

International audience; A longstanding goal in regenerative medicine is to reconstitute functional tissues or organs after injury or disease. Attention has focused on the identification and relative contribution of tissue specific stem cells to the regeneration process. Relatively little is known about how the physiological process is regulated by other tissue constituents. Numerous injury models are used to investigate tissue regeneration, however, these models are often poorly understood. Specifically, for skeletal muscle regeneration several models are reported in the literature, yet the relative impact on muscle physiology and the distinct cells types have not been extensively characterised. We have used transgenic Tg:Pax7nGFP and Flk1GFP/+ mouse models to respectively count the number of muscle stem (satellite) cells (SC) and number/shape of vessels by confocal microscopy. We performed histological and immunostainings to assess the differences in the key regeneration steps. Infiltration of immune cells, chemokines and cytokines production was assessed in vivo by Luminex®.We compared the 4 most commonly used injury models i.e. freeze injury (FI), barium chloride (BaCl2), notexin (NTX) and cardiotoxin (CTX). The FI was the most damaging. In this model, up to 96% of the SCs are destroyed with their surrounding environment (basal lamina and vasculature) leaving a "dead zone" devoid of viable cells. The regeneration process itself is fulfilled in all 4 models with virtually no fibrosis 28 days post-injury, except in the FI model. Inflammatory cells return to basal levels in the CTX, BaCl2 but still significantly high 1-month post-injury in the FI and NTX models. Interestingly the number of SC returned to normal only in the FI, 1-month post-injury, with SCs that are still cycling up to 3-months after the induction of the injury in the other models.We compared the 4 most commonly used injury models i.e. freeze injury (FI), barium chloride (BaCl2), notexin (NTX) and cardiotoxin (CTX). The FI was the most damaging. In this model, up to 96% of the SCs are destroyed with their surrounding environment (basal lamina and vasculature) leaving a "dead zone" devoid of viable cells. The regeneration process itself is fulfilled in all 4 models with virtually no fibrosis 28 days post-injury, except in the FI model. Inflammatory cells return to basal levels in the CTX, BaCl2 but still significantly high 1-month post-injury in the FI and NTX models. Interestingly the number of SC returned to normal only in the FI, 1-month post-injury, with SCs that are still cycling up to 3-months after the induction of the injury in the other models.We compared the 4 most commonly used injury models i.e. freeze injury (FI), barium chloride (BaCl2), notexin (NTX) and cardiotoxin (CTX). The FI was the most damaging. In this model, up to 96% of the SCs are destroyed with their surrounding environment (basal lamina and vasculature) leaving a "dead zone" devoid of viable cells. The regeneration process itself is fulfilled in all 4 models with virtually no fibrosis 28 days post-injury, except in the FI model. Inflammatory cells return to basal levels in the CTX, BaCl2 but still significantly high 1-month post-injury in the FI and NTX models. Interestingly the number of SC returned to normal only in the FI, 1-month post-injury, with SCs that are still cycling up to 3-months after the induction of the injury in the other models.

Published
2016

2. Production of neurospheres from mammalian Müller cells in culture

Author

Claudine Versaux-Botteri, David Hicks, Germaine Michel, Julie Monnin, Nadège Morand-Villeneuve, Laboratoire de Neurosciences Intégratives et Cliniques - UFC (EA 481) (NEURO), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Franche-Comté (UFC), Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Ingénierie et biologie cellulaire et tissulaire (IBCT (ex IFR133)), Centre Hospitalier Régional Universitaire [Besançon] (CHRU Besançon)-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Université de Franche-Comté (UFC), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Université Paris Descartes - Paris 5 (UPD5)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (UR 481) (NEURO), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (EA 481) ( NEURO ), Université Bourgogne Franche-Comté [COMUE] ( UBFC ) -Université de Franche-Comté ( UFC ), Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Ingénierie et biologie cellulaire et tissulaire ( IBCT (ex IFR133) ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Laboratoire de Neurosciences Intégratives et Cliniques - UFC ( NEURO ), and Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC )

Subjects
MESH: Epidermal Growth Factor, MESH: Animals, Newborn, chemistry.chemical_compound, 0302 clinical medicine, MESH: Glial Fibrillary Acidic Protein, MESH: Animals, MESH : Rats, Long-Evans, Cells, Cultured, MESH : Epidermal Growth Factor, 0303 health sciences, Glial fibrillary acidic protein, biology, MESH : Rats, MESH : Animals, Newborn, MESH : Neuroglia, MESH: Retina, Stem Cells, General Neuroscience, MESH : Glial Fibrillary Acidic Protein, Cell biology, medicine.anatomical_structure, MESH : Stem Cells, Neuroglia, MESH: Neuroglia, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Carrier Proteins, MESH: Cells, Cultured, MESH: Rats, MESH: Carrier Proteins, MESH: Stem Cells, Retina, 03 medical and health sciences, MESH: Rats, Long-Evans, Neurosphere, MESH : Cells, Cultured, Glial Fibrillary Acidic Protein, medicine, Animals, Rats, Long-Evans, Progenitor cell, 030304 developmental biology, Epidermal Growth Factor, MESH : Retina, Retinal, Nestin, In vitro, Culture Media, Rats, Animals, Newborn, chemistry, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Culture Media, biology.protein, MESH : Culture Media, MESH : Animals, sense organs, Carrier Proteins, Neuroscience, 030217 neurology & neurosurgery
Abstract

International audience; In lower vertebrates, like fishes and amphibians, retina is able to self-regenerate whereas Mammalian retina has lost this property. Nevertheless, recently, it has turned out that retinal glial Müller cells were playing a role in neuronal regeneration of the adult rodent retina, in case of acute damages, by dedifferentiating and redifferentiating in glial and neural cells. The purpose of this study was to analyse the ability of mammalian Müller cells for forming neurospheres. First of all, rats Müller cells were isolated in a primary culture. Second, these cells were resuspended in two different culture media: the cells cultures in the Neurobasal-A medium kept a typical Müller cells morphology even after 15 days of EGF treatment, and the cells plated in the DMEM-F12 medium formed neurospheres from the third day in culture. The neurosphere cells expressed nestin, cellular retinaldehyde binding protein (CRALBP) and glial fibrillary acidic protein (GFAP). These results showed our capacity to isolate and propagate Müller cells-derived progenitor cells. Moreover, it allows us to control the number of progenitor cells and, in the future, to study their differentiation capacity.

Published
2007

3. Highly efficient SiRNA and gene transfer into hepatocyte-like HepaRG cells and primary human hepatocytes: new means for drug metabolism and toxicity studies

Author

Laurent, Véronique, Glaise, Denise, Nübel, Tobias, Gilot, David, Corlu, Anne, LOYER, Pascal, Génétique, génomique fonctionnelle et biotechnologies (UMR 1078), EFS-Université de Brest ( UBO ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Foie, métabolismes et cancer, Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut de Génétique et Développement de Rennes ( IGDR ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -IFR140-Centre National de la Recherche Scientifique ( CNRS ), Génétique, génomique fonctionnelle et biotechnologies (UMR 1078) (GGB), Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Université de Brest (UBO)-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut de Génétique et Développement de Rennes (IGDR), Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), EFS-Université de Brest (UBO)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Cell Differentiation, MESH : Cell Culture Techniques, Drug-Related Side Effects and Adverse Reactions, Human hepatocytes, MESH: Pharmaceutical Preparations, Cell Culture Techniques, MESH: Stem Cells, MESH : Hepatocytes, Transfection, MESH : RNA, Small Interfering, MESH: Drug Toxicity, MESH : Indicators and Reagents, MESH: Hepatocytes, MESH : Pharmaceutical Preparations, MESH: Cell Proliferation, MESH: RNA, Small Interfering, MESH : Cell Proliferation, Humans, MESH: Electroporation, RNA, Small Interfering, Cell Proliferation, MESH: Cell Culture Techniques, Nucleofection™, Drug metabolism, MESH: Humans, Stem Cells, MESH: Transfection, MESH : Humans, MESH: Indicators and Reagents, MESH : Drug Toxicity, Cell Differentiation, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, In vitro toxicity, Electroporation, Pharmaceutical Preparations, HepaRG cells, Hepatocytes, MESH : Stem Cells, MESH : Electroporation, Indicators and Reagents, [ SDV.MHEP.HEG ] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, MESH : Transfection, MESH : Cell Differentiation
Abstract

International audience; The metabolically competent hepatocyte-like human HepaRG cells represent a suitable alternative in vitro cell model to human primary hepatocytes. Here, we describe the culture procedure required to expand progenitor HepaRG cells and to differentiate them into hepatocyte-like cells. Transient transfection of gene and siRNA into cultured cells, using nonviral strategies, is an invaluable technique to decipher gene functions. In this chapter, we detail transfection protocols for efficient transfer of plasmid DNA or siRNAs into proliferating progenitor or quiescent differentiated HepaRG cells as well as into primary hepatocytes.

Published
2013

4. A bromodeoxyuridine (BrdU) based protocol for characterizing proliferating progenitors in Xenopus embryos

Author

Auger , Hélène, Thuret , Raphaël, El Yakoubi , Warif, Papalopulu , Nancy, Institut de Neurobiologie Alfred Fessard ( INAF ), Centre National de la Recherche Scientifique ( CNRS ), and University of Manchester [Manchester]

Subjects
MESH : Xenopus, MESH : In Situ Hybridization, MESH : Fluorescent Antibody Technique, Indirect, MESH : Antibodies, MESH : Indicators and Reagents, MESH : Embryo, Nonmammalian, MESH : Bromodeoxyuridine, MESH : Microinjections, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Stem Cells, MESH : Cell Proliferation, MESH : Microtomy, MESH : Animals, MESH : S Phase Cell Cycle Checkpoints, MESH : DNA Replication
Abstract

International audience; BrdU is a thymidine analog that is incorporated into DNA during the S-phase of the cell cycle. BrdU incorporation can be used to quantify the number of cells that are in S-phase in the time period that BrdU is available. Thus, BrdU incorporation is an essential method in the quantitative analysis of cell proliferation, during normal embryonic development or after experimental manipulation. It is a reliable and versatile method that can be easily combined with immunohistochemistry and in situ hybridization to relate cell proliferation with gene expression. BrdU incorporation has been used in all model organisms; here, we describe a protocol adapted for use in Xenopus embryos.

Published
2012

5. Aldosterone-producing adenoma formation in the adrenal cortex involves expression of stem/progenitor cell markers

Author

Enzo Lalli, Mathilde Sibony, Xavier Jeunemaitre, Sheerazed Boulkroun, José-Felipe Golib-Dzib, Arndt Benecke, Pierre-François Plouin, Tchao Meatchi, Benoit Samson-Couterie, Maria-Christina Zennaro, Laurence Amar, Estelle Louiset, Hervé Lefebvre, Paris-Centre de Recherche Cardiovasculaire (PARCC - UMR-S U970), Université Paris Descartes - Paris 5 (UPD5)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), Institut des Hautes Etudes Scientifiques (IHES), IHES, Hôpital Européen Georges Pompidou [APHP] (HEGP), Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), CHU Tenon [APHP], Université Pierre et Marie Curie - Paris 6 (UPMC), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), UNIROUEN - UFR Santé (UNIROUEN UFR Santé), Normandie Université (NU)-Normandie Université (NU), Institut de Recherche Interdisciplinaire [Villeneuve d'Ascq] (IRI), Université de Lille, Sciences et Technologies-Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique (CNRS), Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 2019) (UNS), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), Université Côte d'Azur (UCA), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), CHU Tenon [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Droit et Santé-Université de Lille, Sciences et Technologies, Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), Paris-Centre de Recherche Cardiovasculaire ( PARCC - U970 ), Hôpital Européen Georges Pompidou [APHP] ( HEGP ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Paris Descartes - Paris 5 ( UPD5 ), Institut des Hautes Etudes Scientifiques ( IHES ), Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Université Pierre et Marie Curie - Paris 6 ( UPMC ), Différenciation et communication neuronale et neuroendocrine ( DC2N ), Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), UNIROUEN - UFR Santé, Normandie Université ( NU ) -Normandie Université ( NU ), Institut de Recherche Interdisciplinaire [Villeneuve d'Ascq] ( IRI ), Université de Lille, Sciences et Technologies-Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique ( CNRS ), Institut de pharmacologie moléculaire et cellulaire ( IPMC ), Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS ), Université Côte d'Azur ( UCA ), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut des Hautes Études Scientifiques (IHES), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Université Nice Sophia Antipolis (1965 - 2019) (UNS), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA)

Subjects
Aldosterone synthase, MESH: Hypertension, MESH : Adrenal Cortex, chemistry.chemical_compound, 0302 clinical medicine, Endocrinology, Adrenal Glands, MESH : Cell Proliferation, Sonic hedgehog, MESH: Adrenal Glands, Aldosterone, 0303 health sciences, biology, Adrenal cortex, Stem Cells, MESH : Hypertension, medicine.anatomical_structure, Hypertension, MESH : Stem Cells, Zona Glomerulosa, Stem cell, Adenoma, medicine.medical_specialty, MESH: Zona Glomerulosa, education, 030209 endocrinology & metabolism, MESH: Stem Cells, In Vitro Techniques, MESH : Adrenal Glands, 03 medical and health sciences, MESH: Gene Expression Profiling, Zona fasciculata, [ SDV.MHEP ] Life Sciences [q-bio]/Human health and pathology, Internal medicine, MESH: Cell Proliferation, medicine, Humans, Progenitor cell, Cell Proliferation, 030304 developmental biology, MESH: Adenoma, MESH: Humans, MESH : Aldosterone, MESH : Gene Expression Profiling, Gene Expression Profiling, MESH : Humans, MESH: Biological Markers, MESH: Aldosterone, MESH : Biological Markers, MESH : Adenoma, chemistry, Zona glomerulosa, Adrenal Cortex, biology.protein, MESH: Adrenal Cortex, MESH : Zona Glomerulosa, Biomarkers, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

International audience; Aldosterone producing adenoma (APA) is the most common form of surgically curable hypertension. To further understand mechanisms involved in APA formation, we investigated the expression of molecules linked to adrenal stem/precursor cells [β-catenin, Sonic hedgehog (Shh), CD56], and nuclear receptors that play key roles in adrenocortical development and function steroidogenic factor 1, dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1) in six control adrenal glands and 14 adrenals with APA and compared their expression with that of specific markers of zona glomerulosa (ZG) [CYP11B2, Disabled 2 (Dab2)]. Both Dab2 and CD56 were expressed in ZG. Although Dab2 associates uniquely with differentiated ZG cells and its expression is lost when cells transdifferentiate to zona fasciculata (ZF) cells, CD56 was also expressed in ZF and in aldosterone-producing cell clusters, confirming that these structures possess an intermediate phenotype between ZG and ZF cells. Shh was barely detectable in cells located to the outer part of the ZG in the control adrenal; in contrast, its expression was detected in the entire APA and was dramatically increased in the hyperplastic peritumoral ZG. Transcriptome profiling revealed differential expression of components of Shh signaling pathway in a subgroup of APA. Similarly, Wnt/β-catenin signaling was activated in the majority of APA as well as in the entire peritumoral adrenal cortex; however, no mutation was identified in the CTNNB1 gene that could account for β-catenin activation. Our data suggest that both APA and adjacent ZG present characteristics of stem/precursor cells; the reexpression of genes involved in fetal adrenal development could underlie excessive ZG cell proliferation and APA formation.

Published
2011

6. Sumoylation by Ubc9 Regulates the Stem Cell Compartment and Structure and Function of the Intestinal Epithelium in Mice

Author

Esther Danenberg, Alexandra Andrieux, Karim Nacerddine, Ana Cumano, Ghislaine Hamard, Perrine Bomme, Grégory Jouvion, Anne Dejean, Béatrice Romagnolo, Maud D. Demarque, Nick Barker, Hélène Neyret–Kahn, Hans Clevers, Benoit Terris, Hubrecht Institute for Developmental Biology and Stem Cell Research, Organisation Nucléaire et Oncogenèse / Nuclear Organization and Oncogenesis, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Histotechnologie et Pathologie, Institut Pasteur [Paris], Microscopie Ultrastructurale (Plate-forme), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Plateforme Recombinaison Homologue, Transfert d'Embryons et Cryoconservation [Institut Cochin] (PRHTEC), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (UMR_S567 / UMR 8104), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Service d'Anatomie Pathologique, Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Développement des Lymphocytes, Organisation Nucléaire et Oncogenèse, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Plateforme Recombinaison Homologue, Transfert d'Embryons et Cryoconservation [Institut Cochin] ( PRHTEC ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut Cochin ( UMR_S567 / UMR 8104 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), CHU Cochin [AP-HP], Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
SUMO protein, MESH : Tamoxifen, MESH: Mice, Knockout, Mice, 0302 clinical medicine, Keratin, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH: Animals, Intestinal Mucosa, health care economics and organizations, chemistry.chemical_classification, Mice, Knockout, 0303 health sciences, Stem Cells, MESH: Sumoylation, Gastroenterology, LGR5, MESH : Sumoylation, Intestinal epithelium, Cell biology, MESH : Phenotype, medicine.anatomical_structure, Phenotype, Biochemistry, MESH : Stem Cells, MESH: Intestinal Mucosa, [SDV.IMM]Life Sciences [q-bio]/Immunology, Basal lamina, Stem cell, MESH : Ubiquitin-Conjugating Enzymes, education, MESH: Stem Cells, Biology, MESH: Phenotype, MESH : Intestinal Mucosa, 03 medical and health sciences, MESH : Mice, medicine, Animals, MESH: Mice, 030304 developmental biology, Hepatology, Sumoylation, Small intestine, MESH: Ubiquitin-Conjugating Enzymes, Tamoxifen, chemistry, Ubiquitin-Conjugating Enzymes, Keratin 8, MESH : Mice, Knockout, MESH: Tamoxifen, MESH : Animals, 030217 neurology & neurosurgery
Abstract

Background & Aims Small ubiquitin-like modifiers (SUMOs) are attached to other proteins to regulate their function (sumoylation). We investigated the role of Ubc9, which covalently attaches SUMOs to proteins, in the gastrointestinal tract of adult mice. Methods We investigated the effects of decreased sumoylation in adult mammals by generating mice with an inducible knockout (by injection of 4-hydroxytamoxifen) of the E2 enzyme Ubc9 ( Ubc9fl/-/ROSA26-CreERT2 mice). We analyzed the phenotypes using a range of histologic techniques. Results Loss of Ubc9 from adult mice primarily affected the small intestine. Ubc9fl/−/ROSA26-CreERT2 mice died within 6 days of 4-hydroxytamoxifen injection, losing 20% or less of their body weight and developing severe diarrhea on the second day after injection. Surprisingly, other epithelial tissues appeared to be unaffected at that stage. Decreased sumoylation led to the depletion of the intestinal proliferative compartment and to the rapid disappearance of stem cells. Sumoylation was required to separate the proliferative and differentiated compartments from the crypt and control differentiation and function of the secretory lineage. Sumoylation was required for nucleus positioning and polarized organization of actin in the enterocytes. Loss of sumoylation caused detachment of the enterocytes from the basal lamina, as observed in tissue fragility diseases. We identified the intermediate filament keratin 8 as a SUMO substrate in epithelial cells. Conclusions Sumoylation maintains intestinal stem cells and the architecture, mechanical stability, and function of the intestinal epithelium of mice.

Published
2010

7. Human cord blood-derived hematopoietic and neural-like stem/progenitor cells are attracted by the neurotransmitter GABA

Author

Frédéric Deschaseaux, Pierre Tiberghien, Norbert Balon, Vincent Zangiacomi, Stéphane Maddens, Claudine Versaux-Botteri, Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) ( HOTE GREFFON ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Paris Descartes - Paris 5 (UPD5)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC), and Saas, Philippe

Subjects
MESH: Chemotaxis, MESH : Cell Line, Baclofen, MESH: Neurons, Fluorescent Antibody Technique, Gene Expression, MESH: Flow Cytometry, MESH: gamma-Aminobutyric Acid, MESH: GABA Antagonists, MESH: Hematopoietic Stem Cells, GABA Antagonists, 0302 clinical medicine, MESH: Reverse Transcriptase Polymerase Chain Reaction, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, AC133 Antigen, MESH: Receptors, GABA-B, MESH: Antigens, CD, MESH: Fluorescent Antibody Technique, MESH: Receptors, GABA-A, Cells, Cultured, gamma-Aminobutyric Acid, Neurons, MESH : GABA Antagonists, 0303 health sciences, Reverse Transcriptase Polymerase Chain Reaction, MESH: Peptides, MESH : Peptides, Chemotaxis, Stem Cells, MESH : Reverse Transcriptase Polymerase Chain Reaction, MESH : Bicuculline, Cell Differentiation, Hematology, Fetal Blood, Flow Cytometry, MESH : gamma-Aminobutyric Acid, 3. Good health, Cell biology, Endothelial stem cell, MESH: Baclofen, Haematopoiesis, Cord blood, MESH : Stem Cells, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH : Cell Differentiation, MESH : Colony-Forming Units Assay, Adult stem cell, MESH: Cells, Cultured, MESH: Cell Differentiation, MESH: Gene Expression, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH : Flow Cytometry, MESH: Glycoproteins, MESH: Stem Cells, Biology, Bicuculline, MESH : Neurons, Cell Line, Colony-Forming Units Assay, 03 medical and health sciences, MESH: Bicuculline, Antigens, CD, MESH : Cells, Cultured, MESH : Antigens, CD, MESH : Baclofen, Humans, MESH: Colony-Forming Units Assay, MESH: Fetal Blood, GABA-A Receptor Antagonists, Progenitor cell, Glycoproteins, 030304 developmental biology, Interleukin 3, MESH : Fetal Blood, MESH: Humans, MESH : Hematopoietic Stem Cells, MESH : Humans, MESH : Receptors, GABA-A, MESH : Receptors, GABA-B, Cell Biology, Hematopoietic Stem Cells, Receptors, GABA-A, MESH : Glycoproteins, MESH: Cell Line, MESH : Gene Expression, MESH : Fluorescent Antibody Technique, Receptors, GABA-B, nervous system, MESH : Chemotaxis, Cell culture, Immunology, Peptides, GABA-B Receptor Antagonists, 030217 neurology & neurosurgery, Developmental Biology, Homing (hematopoietic)
Abstract

International audience; Migration of stem/progenitor cells is a crucial event for homing toward tissue where cells need to be renewed. The neurotransmitter gamma-aminobutyric acid (GABA) has been shown to have a crucial role in migration of neuronal stem/progenitor cells. Since human umbilical cord blood (HUCB) contains stem/progenitor cells able to generate either neuronal or hematopoietic cells, we evaluated the effect of GABA on this type of cells. While whole fraction of mononuclear cells expressed GABA(A) and GABA(B) receptor subunits (GABA-R), only GABA(B)R subunits were found to be expressed on immature CD133+ cells. Functional experiments revealed that both cell fractions of HUCB were attracted by a gradient of GABA concentration and furthermore were blocked by specific antagonists of GABA(A)R and GABA(B)R bicuculline and saclofen, respectively. Moreover, through GABA(B)R activation the migrating fraction was highly enriched by both hematopoietic progenitors and cells able to generate neuron- like cells in culture. Therefore, GABA is a potent chemoattractant of HUCB stem/progenitor cells specifically through GABA(B)R activation.

Published
2009

8. iNKT cell development is orchestrated by different branches of TGF-beta signaling

Author

Sylvie Martel, C Garcia, Régine Losson, Jean-Marc Doisne, Farhan S. Cyprian, Nadia Duarte, Kamel Benlagha, Kai-Ping Yan, Jean-Benoît Le Luduec, Laurent Bartholin, Julien C. Marie, Branka Horvat, David F. Vincent, Ruth Rimokh, Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Immunologie, génétique et traitement des maladies métaboliques et du diabète (UMR_S 561), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Oncogénèse et progression tumorale, Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Louis Pasteur - Strasbourg I, Virologie humaine, École normale supérieure - Lyon (ENS Lyon)-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), Immunité infection vaccination (I2V), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de biologie et chimie des protéines [Lyon] (IBCP), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure de Lyon (ENS de Lyon)-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Cochin [AP-HP], Immunologie, génétique et traitement des maladies métaboliques et du diabète ( UMR_S 561 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Paris Descartes - Paris 5 ( UPD5 ), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de génétique et biologie moléculaire et cellulaire ( IGBMC ), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), École normale supérieure - Lyon ( ENS Lyon ) -IFR128-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Immunité infection vaccination ( I2V ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-IFR128-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de biologie et chimie des protéines [Lyon] ( IBCP ), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique ( CNRS ), and Peney, Maité

Subjects
MESH: Signal Transduction, MESH : Transcription Factors, MESH : Receptors, Transforming Growth Factor beta, MESH: Spleen, Cellular differentiation, Apoptosis, MESH: T-Lymphocyte Subsets, Mice, 0302 clinical medicine, T-Lymphocyte Subsets, Transforming Growth Factor beta, MESH: Smad4 Protein, Immunology and Allergy, MESH: Animals, Smad4 Protein, 0303 health sciences, biology, Stem Cells, Cell Differentiation, MESH: Transcription Factors, Natural killer T cell, MESH : Mice, Transgenic, Cell biology, MESH: Interleukin-2 Receptor beta Subunit, MESH : Stem Cells, MESH: Receptors, Transforming Growth Factor beta, Stem cell, Signal transduction, MESH : Cell Differentiation, Signal Transduction, MESH : Spleen, MESH: Cell Differentiation, MESH: T-Box Domain Proteins, MESH: Mice, Transgenic, Immunology, MESH : Cell Lineage, Mice, Transgenic, MESH: Stem Cells, Article, 03 medical and health sciences, MESH : Mice, TGF beta signaling pathway, Animals, Cell Lineage, Transcription factor, MESH: Mice, MESH: Transforming Growth Factor beta, MESH : Interleukin-2 Receptor beta Subunit, 030304 developmental biology, MESH : Signal Transduction, Cell growth, MESH: Apoptosis, MESH : Smad4 Protein, Transforming growth factor beta, MESH: Cell Lineage, MESH : Natural Killer T-Cells, MESH : T-Lymphocyte Subsets, Interleukin-2 Receptor beta Subunit, MESH : T-Box Domain Proteins, MESH : Transforming Growth Factor beta, MESH: Natural Killer T-Cells, biology.protein, Natural Killer T-Cells, MESH : Animals, T-Box Domain Proteins, Receptors, Transforming Growth Factor beta, Spleen, MESH : Apoptosis, Transcription Factors, 030215 immunology
Abstract

International audience; Invariant natural killer T (iNKT) cells constitute a distinct subset of T lymphocytes exhibiting important immune-regulatory functions. Although various steps of their differentiation have been well characterized, the factors controlling their development remain poorly documented. Here, we show that TGF-beta controls the differentiation program of iNKT cells. We demonstrate that TGF-beta signaling carefully and specifically orchestrates several steps of iNKT cell development. In vivo, this multifaceted role of TGF-beta involves the concerted action of different pathways of TGF-beta signaling. Whereas the Tif-1gamma branch controls lineage expansion, the Smad4 branch maintains the maturation stage that is initially repressed by a Tif-1gamma/Smad4-independent branch. Thus, these three different branches of TGF-beta signaling function in concert as complementary effectors, allowing TGF-beta to fine tune the iNKT cell differentiation program.

Published
2009

9. CX(3)CR1(+) CD115(+) CD135(+) common macrophage/DC precursors and the role of CX(3)CR1 in their response to inflammation

Author

Cédric Auffray, Ana Cumano, Grégoire Lauvau, Laura Campisi, Frederic Geissmann, Darin K. Fogg, Thierry Jo Molina, David A. Hume, Céline Trouillet, Brigitte Senechal, Julia Leemput, Karine Bigot, Emilie Narni-Mancinelli, Marc Abitbol, Noah Saederup, Israel F. Charo, Système des phagocytes mononucléés et immunopathologie, Université Paris Descartes - Paris 5 (UPD5), Immunologie des maladies infectieuses allergiques et autoimmunes, Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre for Cellular and Molecular Biology of Inflammation, King‘s College London, Gladstone Institute of Cardiovascular Disease, University of California [San Francisco] (UC San Francisco), University of California (UC)-University of California (UC), Centre d'étude et de recherche thérapeutiques en ophtalmologie, The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Développement des Lymphocytes, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by an European Young Investigator (EURYI) award to F. Geissmann, grants from Institut National de la Sant é et de la Recherche M é dicale (Avenir) and Human Frontier Science Program (CDA) to G. Lauvau, grants from the Agence Nationale de la Recherche (ANR IRAP2005) to F. Geissmann and G. Lauvau, and grants from the Fondation pour la Recherche Medicale to F. Geissmann (Equipe FRM 2006) and to G. Lauvau (FRM equipments fund)., Université Nice Sophia Antipolis (... - 2019) (UNS), University of California [San Francisco] (UCSF), University of California-University of California, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Vougny, Marie-Christine, Université Paris Descartes - Paris 5 ( UPD5 ), Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), University of California [San Francisco] ( UCSF ), and Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM )

Subjects
MESH: Inflammation, Monocytes/cytology, MESH: Spleen, MESH: Monocytes, MESH: Listeria monocytogenes, Monocytes, Mice, 0302 clinical medicine, Cell Movement, Spleen/cytology, MESH : Cell Proliferation, MESH : Cell Movement, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, Macrophage, MESH: Animals, MESH: Cell Movement, MESH : Macrophages, MESH : Cell Survival, 0303 health sciences, MESH: Bone Marrow Cells, Stem Cells, hemic and immune systems, Dendritic Cells/enzymology, Receptors, Chemokine/metabolism, 3. Good health, Cell biology, MESH : Phenotype, MESH: Cell Survival, Tumor necrosis factor alpha, Receptors, Chemokine, MESH : Cell Differentiation, MESH: fms-Like Tyrosine Kinase 3, MESH: Receptor, Macrophage Colony-Stimulating Factor, [SDV.IMM] Life Sciences [q-bio]/Immunology, Immunology, CX3C Chemokine Receptor 1, Bone Marrow Cells/cytology, Tumor Necrosis Factor-alpha/biosynthesis, Spleen, Dendritic Cells/cytology, MESH: Phenotype, Article, 03 medical and health sciences, Receptor, Macrophage Colony-Stimulating Factor/metabolism, MESH : Inflammation, fms-Like Tyrosine Kinase 3/metabolism, Tumor Necrosis Factor-alpha, MESH : Listeria Infections, Macrophages, Monocytes/immunology, Dendritic cell, Dendritic Cells, Listeria monocytogenes, fms-Like Tyrosine Kinase 3, Dendritic Cells/immunology, MESH : Dendritic Cells, MESH: Tumor Necrosis Factor-alpha, Reactive Oxygen Species, MESH : Bone Marrow Cells, Myeloid, MESH : Reactive Oxygen Species, Nitric Oxide Synthase Type II, Macrophages/immunology, Inflammation/enzymology, Immunology and Allergy, Listeriosis, MESH : Tumor Necrosis Factor-alpha, MESH: Dendritic Cells, MESH: Reactive Oxygen Species, Cell Differentiation, Inflammation/immunology, Listeriosis/immunology, medicine.anatomical_structure, Phenotype, MESH : Stem Cells, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Nitric Oxide Synthase Type II, medicine.symptom, Stem cell, MESH: Listeria Infections, Nitric Oxide Synthase Type II/metabolism, MESH : Spleen, MESH: Cell Differentiation, MESH : Receptors, Chemokine, MESH : fms-Like Tyrosine Kinase 3, Cell Survival, Macrophages/enzymology, Inflammation, Bone Marrow Cells, Receptor, Macrophage Colony-Stimulating Factor, MESH: Stem Cells, Biology, MESH : Receptor, Macrophage Colony-Stimulating Factor, Reactive Oxygen Species/metabolism, MESH: Cell Proliferation, MESH : Mice, medicine, Animals, MESH: Mice, 030304 developmental biology, Cell Proliferation, MESH: Macrophages, Macrophages/cytology, MESH : Nitric Oxide Synthase Type II, Receptors, Chemokine/deficiency, MESH : Monocytes, Fms-Like Tyrosine Kinase 3, MESH : Animals, MESH : Listeria monocytogenes, MESH: Receptors, Chemokine, Bone Marrow Cells/immunology, 030215 immunology
Abstract

International audience; CX(3)CR1 expression is associated with the commitment of CSF-1R(+) myeloid precursors to the macrophage/dendritic cell (DC) lineage. However, the relationship of the CSF-1R(+) CX(3)CR1(+) macrophage/DC precursor (MDP) with other DC precursors and the role of CX(3)CR1 in macrophage and DC development remain unclear. We show that MDPs give rise to conventional DCs (cDCs), plasmacytoid DCs (PDCs), and monocytes, including Gr1(+) inflammatory monocytes that differentiate into TipDCs during infection. CX(3)CR1 deficiency selectively impairs the recruitment of blood Gr1(+) monocytes in the spleen after transfer and during acute Listeria monocytogenes infection but does not affect the development of monocytes, cDCs, and PDCs.

Published
2009

10. A mono-allelic bivalent chromatin domain controls tissue-specific imprinting at Grb10

Author

Stormy J. Chamberlain, Jean-Philippe Hugnot, Terry Magnuson, Robert Feil, Lionel A. Sanz, Jean Charles Sabourin, Amandine Henckel, Philippe Arnaud, Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Department of Genetics, University of North Carolina [Chapel Hill] (UNC), University of North Carolina System (UNC)-University of North Carolina System (UNC)-Carolina Center for the Genome Sciences, Physiopathologie et thérapie des déficits sensoriels et moteurs, Université Montpellier 2 - Sciences et Techniques (UM2)-IFR76-Institut National de la Santé et de la Recherche Médicale (INSERM), Hamel, Christian, Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), The University of North Carolina at Chapel Hill-Carolina Center for the Genome Sciences, Université Montpellier 2 - Sciences et Techniques ( UM2 ) -IFR76-Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)

Subjects
Male, Cellular differentiation, GRB10 Adaptor Protein, MESH: Neurons, MESH : Promoter Regions, Genetic, MESH: Mice, Knockout, Histones, Mice, 0302 clinical medicine, MESH : Embryo, Mammalian, MESH : Female, MESH: Animals, Imprinting (psychology), Promoter Regions, Genetic, MESH: CpG Islands, Cells, Cultured, Genetics, Mice, Knockout, Neurons, MESH: Histones, 0303 health sciences, biology, MESH : Chromatin, General Neuroscience, Stem Cells, Polycomb Repressive Complex 2, Brain, MESH : GRB10 Adaptor Protein, Cell Differentiation, MESH : CpG Islands, Chromatin, Histone, MESH: Repressor Proteins, MESH : Stem Cells, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Repressor Proteins, MESH : Cell Differentiation, Bivalent chromatin, MESH: Cells, Cultured, MESH: Cell Differentiation, MESH : Male, MESH : Mice, Inbred C57BL, MESH: Stem Cells, General Biochemistry, Genetics and Molecular Biology, Bivalent (genetics), Article, MESH : Neurons, MESH: Chromatin, 03 medical and health sciences, Genomic Imprinting, MESH: Brain, MESH: Mice, Inbred C57BL, MESH : Mice, MESH : Cells, Cultured, MESH: Promoter Regions, Genetic, Animals, MESH : Histones, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Molecular Biology, Gene, MESH: Mice, Alleles, 030304 developmental biology, General Immunology and Microbiology, MESH: Alleles, MESH: Embryo, Mammalian, MESH : Genomic Imprinting, Embryo, Mammalian, MESH: Male, MESH: Genomic Imprinting, Mice, Inbred C57BL, Repressor Proteins, MESH: GRB10 Adaptor Protein, MESH : Brain, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], biology.protein, MESH : Mice, Knockout, CpG Islands, MESH : Animals, Genomic imprinting, MESH : Alleles, MESH: Female, 030217 neurology & neurosurgery
Abstract

International audience; Genomic imprinting is a developmental mechanism that mediates parent-of-origin-specific expression in a subset of genes. How the tissue specificity of imprinted gene expression is controlled remains poorly understood. As a model to address this question, we studied Grb10, a gene that displays brain-specific expression from the paternal chromosome. Here, we show in the mouse that the paternal promoter region is marked by allelic bivalent chromatin enriched in both H3K4me2 and H3K27me3, from early embryonic stages onwards. This is maintained in all somatic tissues, but brain. The bivalent domain is resolved upon neural commitment, during the developmental window in which paternal expression is activated. Our data indicate that bivalent chromatin, in combination with neuronal factors, controls the paternal expression of Grb10 in brain. This finding highlights a novel mechanism to control tissue-specific imprinting.

Published
2008

11. Cord blood-derived neurons are originated from CD133+/CD34 stem/progenitor cells in a cell-to-cell contact dependent manner

Author

Valérie Lapierre, Norbert Balon, Frédéric Deschaseaux, Claudine Versaux-Botteri, Vincent Zangiacomi, Remy Schlichter, Pierre Tiberghien, Stéphane Maddens, Laboratoire de Neurosciences Intégratives et Cliniques - UFC (EA 481) ( NEURO ), Université Bourgogne Franche-Comté [COMUE] ( UBFC ) -Université de Franche-Comté ( UFC ), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) ( HOTE GREFFON ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Institut des Neurosciences Cellulaires et Intégratives ( INCI ), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique ( CNRS ), Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (UR 481) (NEURO), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC), Institut des Neurosciences Cellulaires et Intégratives (INCI), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (EA 481) (NEURO), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Université Paris Descartes - Paris 5 (UPD5)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Neurosciences Intégratives et Cliniques - UFC ( NEURO ), Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC ( HOTE GREFFON ), and Saas, Philippe

Subjects
MESH : Cell Line, Cellular differentiation, CD34, MESH: Neurons, Antigens, CD34, Cell Communication, 0302 clinical medicine, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, AC133 Antigen, MESH: Antigens, CD, Neurons, 0303 health sciences, MESH: Peptides, MESH : Peptides, Stem Cells, Cell Differentiation, Hematology, Fetal Blood, 3. Good health, Cell biology, Endothelial stem cell, medicine.anatomical_structure, Cord blood, MESH : Antigens, CD34, MESH : Stem Cells, [SDV.IMM]Life Sciences [q-bio]/Immunology, Stem cell, MESH : Cell Differentiation, Adult stem cell, Subcellular Fractions, MESH: Cell Differentiation, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH : Cell Communication, MESH: Glycoproteins, MESH: Stem Cells, Biology, MESH : Subcellular Fractions, MESH : Neurons, Cell Line, 03 medical and health sciences, Antigens, CD, MESH: Cell Communication, medicine, MESH : Antigens, CD, Humans, MESH: Fetal Blood, Progenitor cell, 030304 developmental biology, Glycoproteins, MESH : Fetal Blood, MESH: Humans, MESH : Humans, Cell Biology, MESH: Antigens, CD34, MESH : Glycoproteins, MESH: Cell Line, nervous system, MESH: Subcellular Fractions, Immunology, Neuron, Peptides, 030217 neurology & neurosurgery, Developmental Biology
Abstract

International audience; Previous studies described that neurons could be generated in vitro from human umbilical cord blood cells. However, there are few data concerning their origin. Notably, cells generating neurons are not well characterized. The present study deals with the origin of cord blood cells generating neurons and mechanisms allowing the neuronal differentiation. We studied neuronal markers of both total fractions of cord blood and stem/progenitor cord blood cells before and after selections and cultures. We also compared neuronal commitment of cord blood cells to that observed for the neuronal cell line SK-N-BE(2). Before cultures, neuronal markers are found within the total fraction of cord blood cells. In CD133+ stem/progenitor cell fraction only immature neuronal markers are detected. However, CD133+ cells are unable to give rise to neurons in cultures, whereas this is achieved when total fraction of cord blood cells is used. In fact, mature functional neurons can be generated from CD133+ cells only in cell-to-cell close contact with either CD133- fraction or a neurogenic epithelium. Furthermore, since CD133+ fraction is heterogenous, we used several selections to precisely identify the phenotype of cord blood-derived neuronal stem/progenitor cells. Results reveal that only CD34- cells from CD133+ fraction possess neuronal potential. These data show the phenotype of cord blood neuronal stem/progenitor cells and the crucial role of direct cell-to-cell contact to achieve their commitment. Identifying the neuron supporting factors may be beneficial to the use of cord blood neuronal stem/progenitor cells for regenerative medicine.

Published
2008

12. Two types of circulating endothelial progenitor cells in patients receiving long term therapy by HMG-CoA reductase inhibitors

Author

Joseph-Philippe Etievent, Nicolas Meneveau, Frédéric Deschaseaux, Pierre-Emmanuel Falcoz, Nursen Mersin, Pierre Tiberghien, Zohair Selmani, Alfred Penfornis, Sidney Chocron, C. Kleinclauss, Jean-Pierre Kantelip, Siamak Davani, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Marqueurs pronostiques et facteurs de régulations des pathologies cardiaques et vasculaires - UFC ( EA 3920) (PCVP / CARDIO), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Service de Diabétologie - Endocrinologie [CHRU Besançon], Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Saas, Philippe, Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Marqueurs pronostiques et facteurs de régulations des pathologies cardiaques et vasculaires - UFC ( UR 3920) (PCVP / CARDIO), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC ( HOTE GREFFON ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Marqueurs pronostiques et facteurs de régulations des pathologies cardiaques et vasculaires - UFC ( PCVP / CARDIO ), Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ) -Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ), Service de diabétologie - endocrinologie, and Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Jean Minjoz

Subjects
CD31, Male, CD34, MESH : Aged, MESH: Flow Cytometry, Antigens, CD34, Cell Count, Coronary Disease, 030204 cardiovascular system & hematology, MESH : Cell Count, MESH: Cadherins, 0302 clinical medicine, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH : Female, MESH: Endothelial Cells, MESH: Antigens, CD, Cells, Cultured, MESH: Aged, 0303 health sciences, MESH: Middle Aged, Stem Cells, Middle Aged, Cadherins, Flow Cytometry, 3. Good health, Endothelial stem cell, Platelet Endothelial Cell Adhesion Molecule-1, MESH: Leukocytes, Mononuclear, medicine.anatomical_structure, MESH : Antigens, CD31, MESH : Stem Cells, MESH : Antigens, CD34, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, MESH : Cadherins, Stem cell, MESH : Colony-Forming Units Assay, MESH: Cells, Cultured, MESH : Antigens, CD146, medicine.medical_specialty, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH : Flow Cytometry, MESH : Male, MESH : Vascular Endothelial Growth Factor Receptor-2, MESH: Stem Cells, CD146 Antigen, Biology, Endothelial progenitor cell, Colony-Forming Units Assay, 03 medical and health sciences, Antigens, CD, Internal medicine, MESH : Cells, Cultured, MESH: Antigens, CD146, medicine, MESH : Antigens, CD, MESH: Colony-Forming Units Assay, Humans, MESH : Middle Aged, Progenitor cell, MESH: Hydroxymethylglutaryl-CoA Reductase Inhibitors, 030304 developmental biology, Interleukin 3, Aged, Pharmacology, MESH: Humans, MESH: Vascular Endothelial Growth Factor Receptor-2, MESH : Endothelial Cells, MESH: Cell Count, MESH : Humans, MESH : Hydroxymethylglutaryl-CoA Reductase Inhibitors, Endothelial Cells, MESH: Antigens, CD31, MESH: Antigens, CD34, Vascular Endothelial Growth Factor Receptor-2, MESH: Male, MESH : Leukocytes, Mononuclear, Endocrinology, MESH : Coronary Disease, Leukocytes, Mononuclear, Bone marrow, MESH: Coronary Disease, Hydroxymethylglutaryl-CoA Reductase Inhibitors, MESH: Female
Abstract

International audience; 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are widely used to decrease cholesterol synthesis and are well established to reduce vascular diseases. Recently, it has been proposed that statins mobilize endothelial progenitor cells from bone marrow during the first four weeks, which could help to prevent vascular diseases. However, in humans there are few data concerning the long term effects of statin treatment on these endothelial progenitor cells. We investigated whether endothelial progenitor cells can be detected and characterized in patients receiving long term statin therapy. Mononuclear cells from patients receiving or not receiving statin therapy were assessed for progenitor cell content by flow cytometry and were cultured in specific conditions to determine the number and the type of progenitors. Our results showed there were significantly more CD34(+), CD34(+)/CD144(+) circulating progenitor cells in the statin(pos) group than in the statin(neg) group. In culture two types of endothelial progenitor cells were detected. Early endothelial progenitor cells gave colonies at day 5 comprising elongated cells whereas late endothelial progenitor cells generated cobblestone-like colonies with strong proliferation capacities. The number of circulating early endothelial progenitor cells was significantly higher in the statin(neg) group, while only late endothelial progenitor cells were detected in the statin(pos) group. Moreover, cells from cobblestones clearly had an endothelial phenotype CD31(+), VEGF-R2(+), CD34(+), CD146(+) in contrast to cells from colonies from early endothelial progenitor cells, which were VEGF-R2(low), CD34(-). These results strongly suggest that long term statin treatment specifically maintains late endothelial progenitor cells in circulation with a CD34(+)/CD144(+) phenotype.

Published
2006

13. Differentiation of mouse embryonic stem cells into endothelial cells: genetic selection and potential use in vivo

Author

Gimond , Clotilde, Marchetti , Sandrine, Pagès , Gilles, Institut de signalisation, biologie du développement et cancer (ISBDC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Institut de signalisation, biologie du développement et cancer ( ISBDC ), Université Nice Sophia Antipolis ( UNS ), and Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
MESH: Cell Differentiation, MESH : Cell Culture Techniques, Cell Transplantation, Green Fluorescent Proteins, Cell Culture Techniques, Mice, Nude, [SDV.CAN]Life Sciences [q-bio]/Cancer, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Stem Cells, MESH: Extracellular Matrix, MESH : Green Fluorescent Proteins, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, Mice, MESH: Green Fluorescent Proteins, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Mice, MESH : Cells, Cultured, MESH: Mice, Nude, MESH : Embryo, Mammalian, Animals, Humans, MESH: Animals, MESH: Endothelial Cells, MESH: Mice, Cells, Cultured, MESH: Cell Culture Techniques, MESH: Humans, MESH : Endothelial Cells, Reverse Transcriptase Polymerase Chain Reaction, Stem Cells, MESH : Humans, MESH : Reverse Transcriptase Polymerase Chain Reaction, MESH : Extracellular Matrix, MESH: Embryo, Mammalian, MESH : Mice, Nude, Endothelial Cells, MESH: Antigens, CD31, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Differentiation, Embryo, Mammalian, MESH : Cell Transplantation, Extracellular Matrix, Platelet Endothelial Cell Adhesion Molecule-1, MESH: Cell Transplantation, MESH : Antigens, CD31, MESH : Stem Cells, MESH : Animals, MESH : Cell Differentiation, MESH: Cells, Cultured
Abstract

International audience; Embryonic stem (ES) cells are pluripotent cells derived from blastocyst-stage embryos. They are characterized by their infinite self-renewal capacity and their ability to differentiate into many cell types in vitro as well as in vivo. The present protocol describes culture conditions that allow efficient differentiation of mouse ES cells toward the endothelial lineage, both in two-dimensional cultures and in three-dimensional, multicellular embryoid bodies. We also provide a protocol for establishing recombinant ES cell clones, giving the example of cells expressing green fluorescent protein and an antibiotic resistance gene under the control of an endothelial-specific promoter. These transgenes allow the visualization and the genetic selection of endothelial cells from a heterogeneous population of differentiating ES cells. Potential applications for these differentiation models are given, including the study of the endothelial cell lineage and its derivatives, the testing of molecules involved in angiogenesis, and the potential use of selected endothelial cells in vivo.

Published
2006

14. Expression of Pitx2 in stromal cells is required for normal hematopoiesis

Author

Bernard G. Forget, Nicole Navarro, Anne Dubart-Kupperschmitt, Georges Uzan, Brigitte Izac, Philip J. Gage, Cécile Kerdudo, Aurélie Kieusseian, Philippe E. Mangeot, Jalila Chagraoui, Garcia, Marie, Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Human Genetics, University of Michigan [Ann Arbor], University of Michigan System-University of Michigan System, Medicine/Genetics, Yale School of Medicine [New Haven, Connecticut] (YSM), Ontogenese de l'Hematopoiese, Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Yale University School of Medicine, Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Yale School of Medicine, and Université Paris-Sud - Paris 11 (UP11) - Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH : Transcription Factors, MESH: Hematopoiesis, MESH : RNA, Small Interfering, Biochemistry, MESH: Mice, Knockout, Mice, 0302 clinical medicine, MESH: RNA, Small Interfering, MESH: Gene Expression Regulation, Developmental, MESH: Animals, MESH : Homeodomain Proteins, RNA, Small Interfering, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Cells, Cultured, MESH: Lentivirus, Mice, Knockout, 0303 health sciences, PITX2, Stem Cells, Homozygote, Gene Expression Regulation, Developmental, Nuclear Proteins, Hematology, MESH: Transcription Factors, MESH : Fetus, Cell biology, Haematopoiesis, medicine.anatomical_structure, MESH : Lentivirus, Liver, 030220 oncology & carcinogenesis, MESH : Stem Cells, MESH : Transfection, MESH : Colony-Forming Units Assay, MESH: Cells, Cultured, MESH: Homozygote, congenital, hereditary, and neonatal diseases and abnormalities, Stromal cell, Ratón, Immunology, MESH : Mice, Inbred C57BL, MESH: Stem Cells, Biology, Transfection, Colony-Forming Units Assay, 03 medical and health sciences, Fetus, MESH : Homozygote, stomatognathic system, MESH: Mice, Inbred C57BL, MESH : Cells, Cultured, MESH : Mice, MESH: Homeodomain Proteins, Lymph node stromal cell, medicine, Animals, MESH: Colony-Forming Units Assay, Progenitor cell, MESH: Mice, 030304 developmental biology, Homeodomain Proteins, MESH: Transfection, Lentivirus, MESH : Nuclear Proteins, MESH : Liver, MESH: Fetus, Cell Biology, Hematopoiesis, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Mice, Inbred C57BL, MESH : Hematopoiesis, stomatognathic diseases, [INFO.INFO-BT] Computer Science [cs]/Biotechnology, MESH : Stromal Cells, MESH : Mice, Knockout, Bone marrow, MESH : Animals, MESH : Gene Expression Regulation, Developmental, sense organs, Stromal Cells, MESH: Stromal Cells, MESH: Nuclear Proteins, Transcription Factors, MESH: Liver
Abstract

Although the expression of Pitx2, a bicoid family homeodomain transcription factor, is highly regulated during hematopoiesis, its function during this process was not documented; we thus studied hematopoiesis in Pitx2-null mice. We found that Pitx2–/– embryos display hypoplastic livers with reduced numbers of hematopoietic cells, but these cells had normal hematopoietic potential, as evidenced by colony-forming assays, immature progenitor cell assays, and long-term repopulation assays. Because the microenvironment is also crucial to the development of normal hematopoiesis, we established Pitx2–/– and Pitx2+/+ stromas from fetal liver and studied their hematopoietic supportive capacity. We showed that the frequency of cobblestone area-forming cells was 4-fold decreased when using Pitx2–/– stromal cells compared with Pitx2+/+ stromal cells, whatever the Pitx2 genotype of hematopoietic cells tested in this assay. This defect was rescued by expression of Pitx2 into Pitx2–/– fetal liver stromal cells, demonstrating a major and direct role of Pitx2 in the hematopoietic supportive capacity of fetal liver stroma. Finally, we showed a reduced capacity of MS5 stromal cells expressing Pitx2 RNAi to support human hematopoiesis. Altogether these data showed that Pitx2 has major functions in the hematopoietic supportive capacity of fetal liver and adult bone marrow stromal cells.

Published
2006

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